Labeling Protein Lysates with Bocillin-FL

Cell lysates were prepared by bead beating as in the previous literature (Rahlwes et al., 2017 (link)). Then, 30 μg protein of cell lysate was incubated with 100 pmol Bocillin-FL (Invitrogen) in a total volume of 7.5 μL for 30 min in 37°C (Levine and Beatty, 2021 (link)). The sample was mixed with 2.5 μL of 4×-SDS-loading buffer and boiled for 3 min at 98°C. The entire sample was subjected to SDS-PAGE analysis. Gel was imaged by Amersham ImageQuant 800 system (Cytiva).

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Kado T., Akbary Z., Motooka D., Sparks I.L., Melzer E.S., Nakamura S., Rojas E.R., Morita Y.S, & Siegrist M.S. (2023). A cell wall synthase accelerates plasma membrane partitioning in mycobacteria. eLife, 12, e81924.

Publication 2023

Bocillin-FL Amersham ImageQuant 800 system

Bocillin fl Buffer Cell Protein Sds page

Corresponding Organization :

Other organizations : University of Massachusetts Amherst, New York University, Osaka University

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Variable analysis

independent variables

Bocillin-FL concentration (100 pmol)

dependent variables

Protein binding to Bocillin-FL (assessed by SDS-PAGE and imaging)

control variables

Cell lysate protein amount (30 μg)
Incubation time (30 min)
Incubation temperature (37°C)

controls

No controls explicitly mentioned

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